Evaluating the Wind-Induced Mechanical Noise on the InSight Seismometers

The SEIS (Seismic Experiment for Interior Structures) instrument onboard the InSight mission to Mars is the critical instrument for determining the interior structure of Mars, the current level of tectonic activity and the meteorite flux. Meeting the performance requirements of the SEIS instrument is vital to successfully achieve these mission objectives. Here we analyse in-situ wind measurements from previous Mars space missions to understand the wind environment that we are likely to encounter on Mars, and then we use an elastic ground deformation model to evaluate the mechanical noise contributions on the SEIS instrument due to the interaction between the Martian winds and the InSight lander. Lander mechanical noise maps that will be used to select the best deployment site for SEIS once the InSight lander arrives on Mars are also presented. We find the lander mechanical noise may be a detectable signal on the InSight seismometers. However, for the baseline SEIS deployment position, the noise is expected to be below the total noise requirement > 97 % of the time and is, therefore, not expected to endanger the InSight mission objectives

Herpes Simplex Virus 2 Virion Host Shutoff Endoribonuclease Activity Is Required To Disrupt Stress Granule Formation

We previously established that cells infected with herpes simplex virus 2 (HSV-2) are disrupted in their ability to form stress granules (SGs) in response to oxidative stress and that this disruption is mediated by virion host shutoff protein (vhs), a virion-associated endoribonuclease. Here, we test the requirement for vhs endoribonuclease activity in disruption of SG formation. We analyzed the ability of HSV-2 vhs carrying the point mutation D215N, which ablates its endoribonuclease activity, to disrupt SG formation in both transfected and infected cells. We present evidence that ablation of vhs endoribonuclease activity results in defects in vhs-mediated disruption of SG formation. Furthermore, we demonstrate that preformed SGs can be disassembled by HSV-2 infection in a manner that requires vhs endoribonuclease activity and that, befitting this ability to promote SG disassembly, vhs is able to localize to SGs. Together these data indicate that endoribonuclease activity must be maintained in order for vhs to disrupt SG formation. We propose a model whereby vhs-mediated destruction of SG mRNA promotes SG disassembly and may also prevent SG assembly. IMPORTANCE Stress granules (SGs) are transient cytoplasmic structures that form when a cell is exposed to stress. SGs are emerging as potential barriers to viral infection, necessitating a more thorough understanding of their basic biology. We identified virion host shutoff protein (vhs) as a herpes simplex virus 2 (HSV-2) protein capable of disrupting SG formation. As mRNA is a central component of SGs and the best-characterized activity of vhs is as an endoribonuclease specific for mRNA in vivo, we investigated the requirement for vhs endoribonuclease activity in disruption of SG formation. Our studies demonstrate that endoribonuclease activity is required for vhs to disrupt SG formation and, more specifically, that SG disassembly can be driven by vhs endoribonuclease activity. Notably, during the course of these studies we discovered that there is an ordered departure of SG components during their disassembly and, furthermore, that vhs itself has the capacity to localize to SGs

Use of a dual reporter plasmid to demonstrate bactofection with an attenuated aroa- derivative of Pasteurella multocida b:2

A reporter plasmid pSRG has been developed which expresses red fluorescent protein (RFP) from a constitutive prokaryotic promoter within Pasteurella multocida B:2 and green fluorescent protein (GFP) from a constitutive eukaryotic promoter within mammalian cells. This construct has been used to determine the location and viability of the bacteria when moving from the extracellular environment into the intracellular compartment of mammalian cells. Invasion assays with embryonic bovine lung (EBL) cells and an attenuated AroA- derivative of Pasteurella multocida B:2 (strain JRMT12), harbouring the plasmid pSRG, showed that RFP-expressing bacteria could be detected intracellularly at 3 h post-invasion. At this stage, some EBL cells harbouring RFP-expressing bacteria were observed to express GFP simultaneously, indicating release of the plasmid into the intracellular environment. At 5 h post-invasion, more EBL cells were expressing GFP, while still harbouring RFP-expressing bacteria. Concurrently, some EBL cells were shown to express only GFP, indicating loss of viable bacteria within these cells. These experiments proved the functionality of the pSRG dual reporter system and the potential of P. multocida B:2 JRMT12 for bactofection and delivery of a DNA vaccine

The Seismic Experiment for Interior Structure (SEIS): Experiment Data Distribution

The six sensors of SEIS (The Seismic Experiment for Interior Structure) [- one of three primary instruments on NASA's Mars Lander Insight] cover a broad range of the seismic bandwidth, from 0.01 hertz to 50 hertz, with possible extension to longer periods. Data are transmitted in the form of three continuous VBB (Very Broad-Band) components at 2 samples per second (sps), an estimation of the short period (SP) energy content from the SP at 1 sps, and a continuous compound VBB/SP vertical axis at 10 sps. The continuous streams are augmented by requested event data with sample rates from 20 to 100 sps. SEIS data products are downlinked from the spacecraft in raw CCSDS (Consultative Committee for Space Data Systems) packets and converted to both the Standard for the Exchange of Earthquake Data (SEED) format files and ASCII tables (GeoCSV) for analysis and archiving. Metadata are available in dataless SEED and StionXML. Time series data (waveforms) are available in miniseed and GeoCSV. Data are distributed according to FDSN (Federation of Digital Seismograph Networks - http://www.fdsn.org) formats and interfaces. Wind, pressure and temperature data from the Auxiliary Payload Sensor Suite (APSS) will also be available in SEED format, and can be used for decorrelation and diagnostic purposes on SEIS

Constraining Martian Regolith and Vortex Parameters From Combined Seismic and Meteorological Measurements

The InSight mission landed on Mars in November 2018 and has since observed multiple convective vortices with both the high performance barometer and the low-noise seismometer SEIS that has unprecedented sensitivity. Here, we present a new method that uses the simultaneous pressure and seismic measurements of convective vortices to place constraints on the elastic properties of the Martian subsurface and the Martian vortex properties, while also allowing a reconstruction of the convective vortex trajectories. From data filtered in the (0.02–0.3 Hz) frequency band, we estimate that the mean value of η (η = E/[1 − ν2], where E is the Young's modulus and ν is the Poisson's ratio) of the Martian ground in the region around SEIS is 239 ± 140 MPa. In addition, we suggest that the previously reported paucity of vortex seismic observations to the west of InSight may be due to the fact that the ground is harder to the west than to the east, consistent with geomorphological surface interpretations

A Study of Daytime Convective Vortices and Turbulence in the Martian Planetary Boundary Layer Based on Half‐a‐Year of InSight Atmospheric Measurements and Large‐Eddy Simulations

Studying the atmospheric planetary boundary layer (PBL) is crucial to understand the climate of a planet. The meteorological measurements by the instruments onboard InSight at a latitude of 4.5°N make a unique rich data set to study the active turbulent dynamics of the daytime PBL on Mars. Here we use the high‐sensitivity continuous pressure, wind, and temperature measurements in the first 400 sols of InSight operations (from northern late winter to midsummer) to analyze wind gusts, convective cells, and vortices in Mars’ daytime PBL. We compare InSight measurements to turbulence‐resolving large‐eddy simulations (LES). The daytime PBL turbulence at the InSight landing site is very active, with clearly identified signatures of convective cells and a vast population of 6,000 recorded vortex encounters, adequately represented by a power law with a 3.4 exponent. While the daily variability of vortex encounters at InSight can be explained by the statistical nature of turbulence, the seasonal variability is positively correlated with ambient wind speed, which is supported by LES. However, wind gustiness is positively correlated to surface temperature rather than ambient wind speed and sensible heat flux, confirming the radiative control of the daytime Martian PBL; and fewer convective vortices are forming in LES when the background wind is doubled. Thus, the long‐term seasonal variability of vortex encounters at the InSight landing site is mainly controlled by the advection of convective vortices by ambient wind speed. Typical tracks followed by vortices forming in the LES show a similar distribution in direction and length as orbital imagery

Dual Infection and Superinfection Inhibition of Epithelial Skin Cells by Two Alphaherpesviruses Co-Occur in the Natural Host

Hosts can be infected with multiple herpesviruses, known as superinfection; however, superinfection of cells is rare due to the phenomenon known as superinfection inhibition. It is believed that dual infection of cells occurs in nature, based on studies examining genetic exchange between homologous alphaherpesviruses in the host, but to date, this has not been directly shown in a natural model. In this report, gallid herpesvirus 2 (GaHV-2), better known as Marek’s disease virus (MDV), was used in its natural host, the chicken, to determine whether two homologous alphaherpesviruses can infect the same cells in vivo. MDV shares close similarities with the human alphaherpesvirus, varicella zoster virus (VZV), with respect to replication in the skin and exit from the host. Recombinant MDVs were generated that express either the enhanced GFP (eGFP) or monomeric RFP (mRFP) fused to the UL47 (VP13/14) herpesvirus tegument protein. These viruses exhibited no alteration in pathogenic potential and expressed abundant UL47-eGFP or -mRFP in feather follicle epithelial cells in vivo. Using laser scanning confocal microscopy, it was evident that these two similar, but distinguishable, viruses were able to replicate within the same cells of their natural host. Evidence of superinfection inhibition was also observed. These results have important implications for two reasons. First, these results show that during natural infection, both dual infection of cells and superinfection inhibition can co-occur at the cellular level. Secondly, vaccination against MDV with homologous alphaherpesvirus like attenuated GaHV-2, or non-oncogenic GaHV-3 or meleagrid herpesvirus (MeHV-1) has driven the virus to greater virulence and these results implicate the potential for genetic exchange between homologous avian alphaherpesviruses that could drive increased virulence. Because the live attenuated varicella vaccine is currently being administered to children, who in turn could be superinfected by wild-type VZV, this could potentiate recombination events of VZV as well

On‐Deck Seismology: Lessons from InSight for Future Planetary Seismology

Before deploying to the surface of Mars, the short‐period (SP) seismometer of the InSight mission operated on deck for a total of 48 hr. This data set can be used to understand how deck‐mounted seismometers can be used in future landed missions to Mars, Europa, and other planetary bodies. While operating on deck, the SP seismometer showed signals comparable to the Viking‐2 seismometer near 3 Hz where the sensitivity of the Viking instrument peaked. Wind sensitivity showed similar patterns to the Viking instrument, although amplitudes on InSight were ∼80% larger for a given wind velocity. However, during the low‐wind evening hours, the instrument noise levels at frequencies between 0.1 and 1 Hz were comparable to quiet stations on Earth, although deployment to the surface below the Wind and Thermal Shield lowered installation noise by roughly 40 dB in acceleration power. With the observed noise levels and estimated seismicity rates for Mars, detection probability for quakes for a deck‐mounted instrument is low enough that up to years of on‐deck recordings may be necessary to observe an event. Because the noise is dominated by wind acting on the lander, though, deck‐mounted seismometers may be more practical for deployment on airless bodies, and it is important to evaluate the seismicity of the target body and the specific design of the lander. Detection probabilities for operation on Europa reach over 99% for some proposed seismicity models for a similar duration of operation if noise levels are comparable to low‐wind time periods on Mars

A “Coiled-Coil” Motif Is Important for Oligomerization and DNA Binding Properties of Human Cytomegalovirus Protein UL77

Human cytomegalovirus (HCMV) UL77 gene encodes the essential protein UL77, its function is characterized in the present study. Immunoprecipitation identified monomeric and oligomeric pUL77 in HCMV infected cells. Immunostaining of purified virions and subviral fractions showed that pUL77 is a structural protein associated with capsids. In silico analysis revealed the presence of a coiled-coil motif (CCM) at the N-terminus of pUL77. Chemical cross-linking of either wild-type pUL77 or CCM deletion mutant (pUL77ΔCCM) implicated that CCM is critical for oligomerization of pUL77. Furthermore, co-immunoprecipitations of infected and transfected cells demonstrated that pUL77 interacts with the capsid-associated DNA packaging motor components, pUL56 and pUL104, as well as the major capsid protein. The ability of pUL77 to bind dsDNA was shown by an in vitro assay. Binding to certain DNA was further confirmed by an assay using biotinylated 36-, 250-, 500-, 1000-meric dsDNA and 966-meric HCMV-specific dsDNA designed for this study. The binding efficiency (BE) was determined by image processing program defining values above 1.0 as positive. While the BE of the pUL56 binding to the 36-mer bio-pac1 containing a packaging signal was 10.0±0.63, the one for pUL77 was only 0.2±0.03. In contrast to this observation the BE of pUL77 binding to bio-500 bp or bio-1000 bp was 2.2±0.41 and 4.9±0.71, respectively. By using pUL77ΔCCM it was demonstrated that this protein could not bind to dsDNA. These data indicated that pUL77 (i) could form homodimers, (ii) CCM of pUL77 is crucial for oligomerization and (iii) could bind to dsDNA in a sequence independent manner

Novel Small-Molecule Inhibitors of Hepatitis C Virus Entry Block Viral Spread and Promote Viral Clearance in Cell Culture

Combinations of direct-acting anti-virals offer the potential to improve the efficacy, tolerability and duration of the current treatment regimen for hepatitis C virus (HCV) infection. Viral entry represents a distinct therapeutic target that has been validated clinically for a number of pathogenic viruses. To discover novel inhibitors of HCV entry, we conducted a high throughput screen of a proprietary small-molecule compound library using HCV pseudoviral particle (HCVpp) technology. We independently discovered and optimized a series of 1,3,5-triazine compounds that are potent, selective and non-cytotoxic inhibitors of HCV entry. Representative compounds fully suppress both cell-free virus and cell-to-cell spread of HCV in vitro. We demonstrate, for the first time, that long term treatment of an HCV cell culture with a potent entry inhibitor promotes sustained viral clearance in vitro. We have confirmed that a single amino acid variant, V719G, in the transmembrane domain of E2 is sufficient to confer resistance to multiple compounds from the triazine series. Resistance studies were extended by evaluating both the fusogenic properties and growth kinetics of drug-induced and natural amino acid variants in the HCVpp and HCV cell culture assays. Our results indicate that amino acid variations at position 719 incur a significant fitness penalty. Introduction of I719 into a genotype 1b envelope sequence did not affect HCV entry; however, the overall level of HCV replication was reduced compared to the parental genotype 1b/2a HCV strain. Consistent with these findings, I719 represents a significant fraction of the naturally occurring genotype 1b sequences. Importantly, I719, the most relevant natural polymorphism, did not significantly alter the susceptibility of HCV to the triazine compounds. The preclinical properties of these triazine compounds support further investigation of entry inhibitors as a potential novel therapy for HCV infection